EZH2 (Y641N) TR-FRET Assay Kit

The EZH2 (Y641N) TR-FRET Assay Kit is designed to measure activity of the mutant EZH2 complex [EZH2 (Y641N) /EED/SUZ12/RbAp48/AEBP] in a homogeneous 384 reaction format. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The key to the EZH2 (Y641N) Assay Kit is a highly specific antibody that recognizes methylated Histone H3K27. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, Sadenosylmethionine is incubated with a sample containing assay buffer and methyltransferase enzyme for 4 hours. Next, antibody to methylated H3K27 is added. Finally, dye-labeled acceptor is added followed by fluorescence detection.This product has beencited 1 time.

Product Specifications

Background

EZH2 is the catalytic subunit of the PRC2/EED-EZH2 complex, which methylates Lys9 (H3K9me) and Lys27 (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate Lys27 of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4.

UniProt

Q15910

Format

Catalog #ComponentAmountStorage51028EZH2 (Y641N) /EED/SUZ12/ RbAp48/AEBP2100 µg-80°CAvoid freeze/ thaw cycles!5212020 µMS-adenosylmethionine2 x 250 µl-80°CTb-labeled antibody20 µl-80°CBiotinylated histone H3 peptide substrate*500 reactions-80°C52170-A4x HMT Assay Buffer 2A3 ml-20°CDye-labeled acceptor2 x 10 µl-20°CTR-FRET Detection Buffer4 ml-20°C79969White, Nonbinding Corning, low volume, microtiter plate1Room Temp*Resuspend in 500 µl of distilled water

Shipping Conditions

-80°C

Storage Conditions

At least 6 months from date of receipt when stored as directed.

Notes

MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:Fluorescent microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) Adjustable micropipettor and sterile tipsThis mutation is often cited in the scientific literature as Y641N in variant 3 (GenBank Accession No. NM_001203247) .